剩余活性污泥中微生物蛋白质的提取工艺研究

利用剩余活性污泥中含有大量的微生物的特点,从中提取蛋白质是有效减少污泥二次污染的重要途径。运用单因素实验的方法,以蛋白质的"盐提-酸沉"为主要提取思路,同时结合研磨、离心、搅拌等多种工艺手段。改变提取液盐浓度、提取pH、样品和提取液的体积比、提取温度、沉淀pH、沉淀温度等参数对活性污泥中提取蛋白质效果的影响进行了研究。     

紫色光合细菌捕获太阳能的分子机理

光合作用是地球上最重要的化学反应,生物体通过它捕获太阳能,转为化学能供生长繁殖需要.光合细菌是地球上最早出现的具有原始光能合成体系的微生物,其光合反应中心是一个由多种色素分子与蛋白质以非共价键方式结合的、具有特定构象的色素-蛋白复合体-光反应中心RC(Reaction center)和LH(Light Harvesting),光能通过电荷分离及电子转移反应转化为化学能,其效率是当前人工模拟远远不能及的.本文综述了紫色光合细菌捕获太阳能的分子结构、作用机理的研究进展,并结合作者在R.sphaeroides LHII蛋白组份同源及异源基因表达方面的研究结果进行相应的分析,明确了Rhodobacter sphaeroides基因组中同源基因puc2BA的表达特点和功能,Rhodovulum sulfidophilum pucsBA与R.sphaeroides pufBA能够同时在R.sphaeroides中表达,能同时形成LHII和LHI,并具有能量传递功能.     

非生物胁迫对盐生杜氏藻DsMPK转录、翻译和磷酸化修饰的影响

促有丝分裂活化蛋白激酶(Mitogen-activated protein kinase,MAPK或MPK)途径是在真核生物中广泛存在的调控多种生理过程的信号途径,在细胞增殖、分化、凋亡、生物和非生物胁迫等生理过程的调控中起着关键的作用.盐生杜氏藻(Dunaliella salina,盐藻)是目前世界上最耐盐的真核光合生物之一.在前期实验中,已发现盐藻MAPK(DsMPK)的表达受到低温和高盐胁迫的抑制.分析DsMPK在高温、低盐、氧化、紫外胁迫中的表达变化,发现DsMPK还受到氧化胁迫的抑制.进而分析DsMPK在高温、低盐、氧化、低温、高盐和紫外胁迫中翻译和磷酸化修饰的变化,发现除低盐胁迫外,在磷酸化修饰水平DsMPK同样受到各种非生物胁迫的抑制.在对不同生长阶段盐藻DsMPK磷酸化变化的分析中还发现,DsMPK的磷酸化水平与生长速率有一致性.各方面DsMPK的变化趋势说明其为一调控生长相关的MAPK.     

酶联免疫吸附法测定麻疯树核糖体失活蛋白

建立了一种检测麻疯树核糖体失活蛋白(Curcin)的简便、快速的酶联免疫吸附测定法(ELISA).ELISA条件优化试验结果显示:最佳条件为costar酶标板在紫外线下照射45 min后进行包被;抗原最适稀释度为1:640倍(2.5μg/mL);curcin抗血清最适稀释度为1:12 800倍;2%封闭用山羊血清封闭120 min;二抗最适稀释度为1:5 000倍.在此试验条件下检测curcin含量,得到回归方程y=31.722x+106.88,相关系数R2=0.993 3.Curcin浓度在0.08～11μg/mL之间,曲线呈良好的线性关系.本方法的最低检测限为0.08μg/mL.高、中、低3个浓度的添加回收率分别为95.56%、102.27%、98.40%,平均回收率为98.74%.板内和板间的平均变异系数分别为4.78%和6.71%.Curcin抗血清与2种大戟科同类蛋白均无交叉反应.利用本方法成功检测了麻疯树种仁及脱毒饼粕中的curcin含量.     

镉对瓯江彩鲤脏器组织蛋白质的氧化损伤作用

瓯江彩鲤(Cyprinus carpio var color)是浙江省大力推广的主要稻田养鱼鱼种,重金属污染对其生态毒理效应研究具有重要意义.针对镉胁迫瓯江彩鲤蛋白质氧化损伤效应,采用静态置换法,研究了0.412～2.059 mg·L-1浓度范围内的镉对瓯江彩鲤鳃、脑和肝脏组织蛋白质的氧化损伤作用.结果表明,瓯江彩鲤鳃...     

Colonization of Penicillium oxalicum SL2 in Pb-contaminated paddy soil and its immobilization effect on soil Pb

Penicillium oxalicum SL2 (SL2) is a previously screened Pb-tolerant fungus that can promote crops growth. The relationship between SL2 colonization and Pb immobilization was studied to provide a theoretical basis for microbial remediation of Pb-contaminated paddy soil. In this study, green fluorescent protein (GFP) labeled SL2 was inoculated into different Pb-contaminated paddy soils (S1-S6). The Pb extracted from the soil by HNO₃, EDTA and CaCl₂ were used to characterize the available Pb. The results showed that the colonization of SL2 was divided into lag phase (0-7 days), growth phase (7-30 days), and mortality phase (30-90 days). SL2 colonized well in sandy soils rich in clay and total phosphorus with initial pH of 4.5-7.0. In addition, SL2 increased soil pH and decreased soil Eh, which was beneficial to immobilize Pb. In different soils, the highest percentages of CaCl₂-Pb, EDTA-Pb, and HNO₃-Pb immobilized by SL2 were 34.34%-40.53%, 17.05%-20.11%, and 7.39%-15.62%, respectively. Pearson correlation analysis showed that the percentages of CaCl₂-Pb and EDTA-Pb immobilized by SL2 were significantly positively correlated with the number of SL2 during the growth phase. SL2 mainly immobilized Pb in the growth phase and a higher peak number of SL2 was beneficial to the immobilization of Pb.     

Alginate modifies the physiological impact of CeO2 nanoparticles in corn seedlings cultivated in soil

Alginates are naturally occurring components of organic matter in natural soil whose effects on nanoparticle （NP） toxicity to plants is not well understood. In the present study, corn plants were grown for one month in soil spiked with 400 mg/kg CeO2 NPs with various alginate concentrations. After one month of growth in the NPs impacted soil, plants were harvested and analyzed for Ce and mineral element concentrations. Chlorophyll concentration and heat shock protein 70, used as biomarkers for oxidative stress, were also evaluated. Results showed that, compared to CeO2 NPs treatment, alginate at 10, 50, and 100 mg/kg increased Ce concentration in roots by approximately 46%, 38%, and 29% and by 115%, 45%, and 56% in shoots, respectively. CeO 2 NPs without alginate increased Mn accumulation in roots by 34% compared to control. CeO2 NPs with low and medium alginate increased Mn by ca. 92% respect to NPs without alginate and by ca. 155% respect to control. CeO2 NPs without/with alginate significantly increased accumulation of Fe and Al in roots. In addition, alginate at 50 mg/kg increased Zn accumulation in roots by 52% compared to control. In shoots, K increased at all NP treatments but the accumulation of other elements was not affected. Alginate enlarged the impact of CeO2 NPs to corn plants by reducing chlorophyll a content and triggering overexpression of heat shock protein 70.     

蛋白质对PVDF超滤膜污染行为的界面微观作用力解析

采用原子力显微镜,结合自制微颗粒探针,对膜-污染物及污染物-污染物间微观作用力进行了检测分析,考察了不同p H条件下牛血清蛋白(BSA)在不同界面特性PVDF超滤膜上的膜污染行为.结果表明,在膜过滤初期,通量剧烈衰减主要由BSA和膜之间黏附力作用导致,在膜过滤后期,BSA与BSA之间的黏聚作用则是影响后期膜污染行为的主要因素;PA膜-BSA和PP膜-BSA之间的黏附作用力均大于BSA-BSA之间的黏聚作用力,说明在整个膜过滤过程中,BSA与PVDF超滤膜之间的黏附作用对膜污染起主导作用;相同p H条件下,PA膜-BSA之间的微观作用小于PP膜-BSA之间的相互作用力,说明亲水性较强的PA膜具有较强的抗污染性;而PA膜和PP膜过滤后期BSA-BSA之间黏聚力相差不大,也进一步说明膜过滤过程中膜-污染物之间的微观作用力是影响膜污染行为的主要因素.     

pH值对SBR生物反应器出水溶解态有机质含量与构成的影响

为探讨污水二级生化出水中有机物(EfOM)的构成特点和变化规律,采用配置生活污水,对不同pH值运行条件下, SBR生物反应器出水中的溶解态有机质(DOM)含量和构成进行了比较研究.结果表明,运行期间pH值的调节对SBR反应器出水DOM含量与构成的影响均较大.初始pH为6.5的SBR反应器, pH值降低为6.0后,出水DOC由原来的4.0mg·L-1不断升高,进一步提高pH值,能够起到一定的缓解作用;初始pH为8.0的SBR反应器,出水DOC从4.0mg·L-1快速降低并保持在2mg·L-1左右, pH值降低为6.5后, DOC变化不大.单宁酸和蛋白质、碳水化合物、DNA等微生物代谢产物(SMPs)是出水DOM的主要组成部分, pH值对SMPs生成的影响是一个长期的过程,运行过程中降低pH值,造成出水SMPs含量不断升高. pH值对整个循环过程中溶解态有机物降解情况的影响,在厌氧阶段发挥了主要作用,较高pH值有利于厌氧阶段有机物的降解,而好氧阶段的有机物降解受pH值影响不大.     

蛋白质组学技术在环境内分泌干扰物分析上应用

为了探讨蛋白质组学技术在环境雌激素分析上的应用,用浓度为3.7×10-9mol/L的17β-雌二醇(E2)处理MCF-7细胞2或3天后进行双向电泳,对比不同的蛋白质提取、染色方法,利用光密度扫描仪将电泳图像数字化,并使用PDQuest软件进行图像分析,以观察雌激素诱导下的蛋白质表达图谱。结果表明,在蛋白质的提取上,冻融法比超声裂解法更有效;在蛋白质染色上,银染法比考马斯亮蓝染色法能得到更多可显示的蛋白质斑点。用E2处理72h后的MCF-7细胞,可检测到40个下调蛋白和5个上调蛋白。该蛋白质表达模式的改变提示了环境雌激素诱导下的特异性蛋白质表达图谱变化。本研究为分析雌激素类内分泌干扰物的内在作用机制提供了一种新的可行方法,也为建立其高通量筛选方法提供了新的思路。